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. 2021 Jun 15;11(6):887.
doi: 10.3390/biom11060887.

Ethanolic Extracts of Adlay Testa and Hull and Their Active Biomolecules Exert Relaxing Effect on Uterine Muscle Contraction through Blocking Extracellular Calcium Influx in Ex Vivo and In Vivo Studies

Yun-Ju Huang  1 , Yu-Chieh Chen  2 , Hsin-Yuan Chen  1 , Yi-Fen Chiang  1 , Mohamed Ali  3 , Wenchang Chiang  2 , Cheng-Pei Chung  4 , Shih-Min Hsia  1   5   6   7
Affiliations

Ethanolic Extracts of Adlay Testa and Hull and Their Active Biomolecules Exert Relaxing Effect on Uterine Muscle Contraction through Blocking Extracellular Calcium Influx in Ex Vivo and In Vivo Studies

Yun-Ju Huang et al. Biomolecules. .

Abstract

Dysmenorrhea is one of the most prevalent disorders in gynecology. Historically, adlay (Coix lachryma-jobi L. var. Ma-yuen Stapf.) has been explored for its anti-tumor, pain relief, anti-inflammatory, and analgesic effects. The aim of this study was to evaluate the effects of adlay seeds on the inhibition of uterine contraction and thus dysmenorrhea relief, in vitro and in vivo. HPLC-MS and GC were used to elucidate the ethyl acetate fraction of adlay testa ethanolic extract (ATE-EA) and ethyl acetate fraction of adlay hull ethanolic extract (AHE-EA). Elucidation yielded flavonoids, phytosterols, and fatty acids. Uterine leiomyomas and normal adjacent myometrial tissue were evaluated by oxytocin- and PG-induced uterine contractility. ATE-EA and AHE-EA suppressed uterine contraction induced by prostaglandin F2 alpha (PGF2α), oxytocin, carbachol, and high-KCl solution ex vivo. In addition, the external calcium (Ca2+) influx induced contraction, and increased Ca2+ concentration was inhibited by ATE-EA and AHE-EA on the uterine smooth muscle of rats. Furthermore, ATE-EA and AHE-EA effectively attenuated the contraction of normal human myometrium tissues more than adjacent uterine leiomyoma in response to PGF2α. 3,5,6,7,8,3',4'-Heptamethoxyflavone and chrysoeriol produced a remarkable inhibition with values of IC50 = 24.91 and 25.59 µM, respectively. The experimental results showed that treatment with ATE-EA at 30 mg/day effectively decreased the writhing frequency both on the oxytocin-induced writhing test and acetic acid writhing test of the ICR mouse.

Keywords: Coix lachryma-jobi; acetic acid writhing; dysmenorrhea; oxytocin-induced writhing; smooth muscle.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of adlay on the uterine contraction induced by PGF2α (10−6 M). (A) Effect of different parts of adlay on isolated rat’s uterine contractions. (B) Inhibitory effect of AHE subfractions on isolated rat’s uterine contraction. (C) Inhibitory effect of ATE subfractions on isolated rat’s uterine contraction. Data represent mean values ± SD. * p < 0.05, ** p < 0.01 compared with control. AHE, ethanolic extracts of adlay hull; ATE, ethanolic extracts of adlay testa; ABE, ethanolic extracts of adlay bran; PAE, ethanolic extracts of polished adlay; AHE-Wa, H2O fraction of adlay hull ethanolic extract; AHE-Bu, 1-butanol fraction of adlay hull ethanolic extract; AHE-EA, ethyl acetate fraction of adlay hull ethanolic extract; AHE-Hex, n-hexane fraction of adlay hull ethanolic extract; ATE-Wa, H2O fraction of adlay testa ethanolic extract; ATE-Bu, 1-butanol fraction of adlay testa ethanolic extract; ATE-EA, ethyl acetate fraction of adlay testa ethanolic extract; ATE-Hex, n-hexane fraction of adlay testa ethanolic extract.
Figure 2
Figure 2
The effect of different concentrations of ATE-EA on uterine contraction amplitude under influence of PGF2α (A), oxytocin (B), carbachol (C), and KCl solution (D). The effect of different concentrations of AHE-EA on uterine contraction amplitude under influence of PGF2α (E), oxytocin (F), carbachol (G), and KCl solution (H). Data represent mean values ± SD. * p < 0.05, ** p < 0.01 compared with control. PGF2α, prostaglandin F2 alpha; ATE-EA, ethyl acetate fraction of adlay testa ethanolic extract; AHE-EA, ethyl acetate fraction of adlay hull ethanolic extract.
Figure 3
Figure 3
Effects of ATE-EA given cumulatively on the PGF2α-induced (A), oxytocin-induced (B), carbachol-induced (C), and KCl solution-induced (D) contractions of isolated rat uterus. AHE-EA given cumulatively on the PGF2α-induced (E), oxytocin-induced (F), carbachol-induced (G), and KCl solution-induced (H) contractions of isolated rat uterus. Data represent mean values ± SD. * p < 0.05, ** p < 0.01 compared with control. PGF2α, prostaglandin F2 alpha; ATE-EA, ethyl acetate fraction of adlay testa ethanolic extract; AHE-EA, ethyl acetate fraction of adlay hull ethanolic extract.
Figure 4
Figure 4
Inhibitory effect of ATE-EA and AHE-EA on calcium (Ca2+)-dependent contractile responses ex vivo. (A,B) Calcium chloride (0.1–10 mM) added into the bathing solution resulted in a concentration-dependent increase in contraction of the isolated rat uterus, which was inhibited by (C) ATE-EA (175 μg/mL) and (D) AHE-EA (175 μg/mL).
Figure 5
Figure 5
Inhibitory effects of serial concentrations of ATE-EA and AHE-EA on contractions of isolated human uterus induced by PGF2α (A,E), oxytocin (B,F), carbachol (C,G), and KCl solution (D,H). Data represent mean values ± SD. ** p < 0.01 compared with control. PGF2α, prostaglandin F2 alpha; ATE-EA, ethyl acetate fraction of adlay testa ethanolic extract; AHE-EA, ethyl acetate fraction of adlay hull ethanolic extract.
Figure 6
Figure 6
Effects of DMSO (A) and serial concentrations of ATE-EA (B) and AHE-EA (C) on PGF2α-induced contraction amplitude on both normal myometrial tissue and leiomyoma tissue, respectively. Data represent mean values ± SD. ** p < 0.01 compared with control. # p < 0.05, ## p < 0.01 compared with normal human uterine tissue. PGF2α, prostaglandin F2 alpha; ATE-EA, ethyl acetate fraction of adlay testa ethanolic extract; AHE -EA, ethyl acetate fraction of adlay hull ethanolic extract.
Figure 7
Figure 7
Total ion chromatogram of extracts from adlay (A) ATE-EA and (B) AHE-EA.
Figure 7
Figure 7
Total ion chromatogram of extracts from adlay (A) ATE-EA and (B) AHE-EA.
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